The Homologous Series Method

Shaltiel's homologous series method of synthesizing hydrocarbon-coated agaroses was supplemented by the so-called exploratory kit, for choosing the most appropriate column and for optimizing resolution. This analytical kit, which was commercially available for some years, contained a homologous series of small columns from Seph-C1 to Seph-C10 with two control columns. The principle was to determine the lowest member of the homologous series capable of retaining the desired enzyme or protein. This column was then selected for the purification of the desired protein. In a second step it was attempted to increase resolution by optimizing the elution procedure which ranged from mild salting-in procedures to reversible denaturation steps. This procedure or variants thereof are still the method of choice for most groups. However as illustrated by Oscarsson et al., the number of failures is probably very high.

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